Zhenyuan Wang

Analysis of postmortem metabolic changes in rat kidney cortex using Fourier transform infrared spectroscopy

Estimation of the time since death (postmortem interval, PMI) is one of the most difficult problems in forensic investigations and many methods currently are utilized to estimate the PMI. The aim of this study was to investigate the changes of Fourier transform infrared (FT-IR) spectra of rat kidney cortex from time zero to 168 h postmortem at molecular level. The spectra of rat kidney cortex displayed the prominent changes with increasing postmortem interval. (1) Significant increase in the intensity of the C-H stretching bands at 2958, 2925, 2871, 2852 cm −1 ,t he=C-H stretching band at 3012 cm −1 , the CO2 − symmetric stretching band at 1396 cm −1 and the N-H bend, C-N stretching at 1541 cm −1 ; (2) significant decrease in the intensity of the PO2 − stretching band at 1238, 1080 cm −1 ; (3) the intensity of at 3303, 1652 and 1170 cm −1 remained relatively stable. The linear regression analysis of the various absorption intensity and area ratios against the PMI shows a close correlation, maximum for A1541/A1396 ratio (R 2 = 0.95) and minimum for I1080/I1396 ratio (R 2 = 0.70). Our results indicate

Attenuated Total Reflection Fourier Transform Infrared Spectroscopic Investigation of the Postmortem Metabolic Process in Rat and Human Kidney Cortex

Attenuated total reflection (ATR) Fourier transform infrared (FT-IR) spectroscopy has been applied to study the short and long term postmortem metabolic processes in rat and human kidney cortexes. The goals of this project were as follows: (1) to investigate the changes of ATR spectra in different rat and human tissues after death, (2) to explore the best mathematical model with different band absorption ratio changes to determine the postmortem interval (PMI), and (3) to establish a preliminary human postmortem ATR spectra database. There were three different types of metabolic changes after death based on the spectral results: (1) the intensities of some bands increased continuously (e. g., C–H stretching region), (2) the intensities of other bands decreased continuously (e.g., PO2− symmetric stretching), and (3) other bands remained relatively stable (e.g., C–OH bending, CO–O–C antisymmetric stretching). The band absorbance ratios for rats were found to display either a significant increase or decrease with increasing time after death. Of the absorbance ratios of the various bands investigated to find the best fit with the cubic model function in rats, the A1652/A1396 ratio showed the strongest correlation (R2 = 0.937). Comparison of the rat kidney cortex spectra with selected human postmortem cases showed similar postmortem metabolic changes. In conclusion, ATR FT-IR spectroscopy was shown to be a convenient and reliable method of determining short and long term postmortem intervals by simultaneously monitoring several specific parameters, although these observations have yet to be applied at forensic scenes by further field studies.

Estimation of Postmortem Interval in Rat Liver and Spleen Using Fourier Transform Infrared Spectroscopy

ABSTRACT The aim of this study was to investigate the changes of Fourier transform infrared (FT-IR) spectra of rat liver and spleen tissues from time zero to 144 h postmortem. The absorbance (Ax represents the infrared absorbance at wavenumber x cm−1) ratios of major absorbance bands were examined. A 3303/A 2925, A 1647/A 1541 (liver), A 1238/A 1338, A 1153/A 1338 (liver), A 1080/A 1338, and A 1030–1050/A 1338 (liver) showed a decrease postmortem, whereas the A 1396/A 1456 ratio (spleen) showed an increase. Furthermore, these absorbance ratios showed a strong linear relationship with increasing time postmortem, and the A 1238/A 1338 ratio of spleen offered a stronger correlation (R 2 = 0.88) than did the others. A new absorbance band appeared at 1120 cm−1 for the liver and spleen at 120 h postmortem and another new band appeared at 1170 cm−1 for the liver, but the existing absorbance band at 1170 cm−1 disappeared in the spleen. Our initial results demonstrate that the quantitative analysis of FT-IR spectrum related to postmortem interval (PMI) shows a strong linear correlation between absorbance ratios and increasing time after death. The FT-IR changes of spectra and absorbance ratios can support further study for estimating short-term and long-term PMI. Upon future validation, FT-IR can offer advantages in combination with established methods to improve PMI estimation.

Characterization of postmortem biochemical changes in rabbit plasma using ATR-FTIR combined with chemometrics: A preliminary study

Postmortem interval (PMI) determination is one of the most challenging tasks in forensic medicine due to a lack of accurate and reliable methods. It is especially difficult for late PMI determination. Although many attempts with various types of body fluids based on chemical methods have been made to solve this problem, few investigations are focused on blood samples. In this study, we employed an attenuated total reflection (ATR)-Fourier transform infrared (FTIR) technique coupled with principle component analysis (PCA) to monitor biochemical changes in rabbit plasma with increasing PMI. Partial least square (PLS) model was used based on the spectral data for PMI prediction in an independent sample set. Our results revealed that postmortem chemical changes in compositions of the plasma were time-dependent, and various components including proteins, lipids and nucleic acids contributed to the discrimination of the samples at different time points. A satisfactory prediction within 48h postmortem was performed by the combined PLS model with a good fitting between actual and predicted PMI of 0.984 and with an error of ±1.92h. In consideration of the simplicity and portability of ATR-FTIR, our preliminary study provides an experimental and theoretical basis for application of this technique in forensic practice.

Research progress in the estimation of the postmortem interval by Chinese forensic scholars

ABSTRACT The determination of time since death or the postmortem interval (PMI) is one of the most important and frequently asked questions in forensic medicine. Medicolegal scholars and forensic pathologists around the world have studied the estimation of PMI extensively in the past, and many novel methods and advanced technologies have now been applied in the field. For several centuries, Chinese forensic examiners have also worked on the estimation of the PMI, and there are a large number of excellent studies published in Chinese rather than in English, and these are not easily accessible or known internationally. Therefore we have conducted a review of relevant studies published by Chinese forensic scholars in the last few decades. The scope of this review is to provide a concise summary of the current progress in the estimation of PMI by Chinese forensic researchers using molecular biology, spectroscopic technology, entomological methods, energy changes, thanatochemistry and other methods.

Estimation of the late postmortem interval using FTIR spectroscopy and chemometrics in human skeletal remains

Due to a lack of reliable and accurate methods, determining the postmortem interval (PMI) of human skeletal remains is one of the most important and challenging tasks in forensic medicine. In this paper, we studied the changes to bone chemistry with increasing PMI in two different experimental conditions using Fourier transform infrared (FTIR) spectroscopy in conjunction with chemometrics methods Paired bone samples collected from 56 human corpses were buried (placed in soil) and unburied (exposed to the air) for intervals between 76 and 552 days. The results of principle component analysis (PCA) showed the chemical differences of these two cases had a significant influence on the rate of decomposition of the remains. Meanwhile, satisfactory predictions were performed by the genetic algorithm combined with partial least-squares (GA-PLS) with the root mean square errors of prediction (RMSEP) of 50.93days for buried bones and 71.03days for unburied bones. Moreover, the amide I region of proteins and the area around 1390cm-1, which is associated with fatty acids, were identified with regular changes by GA-PLS and played an important role in estimating PMI. This study illustrates the feasibility of utilizing FTIR spectroscopy and chemometrics as an attractive alternative for estimating PMI of human remains and the great potential of these techniques in real forensic cases with natural conditions.

Application of Fourier transform infrared spectroscopy with chemometrics on postmortem interval estimation based on pericardial fluids

Postmortem interval (PMI) evaluation remains a challenge in the forensic community due to the lack of efficient methods. Studies have focused on chemical analysis of biofluids for PMI estimation; however, no reports using spectroscopic methods in pericardial fluid (PF) are available. In this study, Fourier transform infrared (FTIR) spectroscopy with attenuated total reflectance (ATR) accessory was applied to collect comprehensive biochemical information from rabbit PF at different PMIs. The PMI-dependent spectral signature was determined by two-dimensional (2D) correlation analysis. The partial least square (PLS) and nu-support vector machine (nu-SVM) models were then established based on the acquired spectral dataset. Spectral variables associated with amide I, amide II, COO−, C-H bending, and C-O or C-OH vibrations arising from proteins, polypeptides, amino acids and carbohydrates, respectively, were susceptible to PMI in 2D correlation analysis. Moreover, the nu-SVM model appeared to achieve a more satisfactory prediction than the PLS model in calibration; the reliability of both models was determined in an external validation set. The study shows the possibility of application of ATR-FTIR methods in postmortem interval estimation using PF samples.

Estimation of the age of human bloodstains under the simulated indoor and outdoor crime scene conditions by ATR-FTIR spectroscopy

Estimation of the age of human bloodstains is of great importance in forensic practices, but it is a challenging task because of the lack of a well-accepted, reliable, and established method. Here, the attenuated total reflection (ATR)-Fourier transform infrared (FTIR) technique combined with advanced chemometric methods was utilized to determine the age of indoor and outdoor bloodstains up to 107 days. The bloodstain storage conditions mimicked crime scene scenarios as closely as possible. Two partial least squares regression models—indoor and outdoor models with 7–85 days—exhibited good performance for external validation, with low values of predictive root mean squared error (5.83 and 4.77) and high R2 values (0.94 and 0.96) and residual predictive deviation (4.08 and 5.14), respectively. Two partial least squares–discriminant analysis classification models were built and demonstrated excellent distinction between fresh (age ≤1 d) and older (age >1 d) bloodstains, which is highly valuable for forensic investigations. These findings demonstrate that ATR-FTIR spectroscopy coupled with advanced chemometric methods can be employed as a rapid and non-destructive tool for age estimation of bloodstains in real-world forensic investigation.

Application of MALDI-TOF MS for Estimating the Postmortem Interval in Rat Muscle Samples†

Estimating the postmortem interval (PMI) is very important in the forensic sciences. Although many approaches have been used for estimating the PMI, accurate PMI calculations are still difficult. In this study, four Sprague Dawley (SD) rats were sacrificed by suffocation, and muscle samples were collected by dissection at various time intervals (0, 48, 96, and 144 h) after death. All samples were probed using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) to obtain molecular images and data for principal component analysis (PCA). The results showed that the peaks at m/z 1511, 1543, 1564, 1586 clearly decreased in intensity from 0 to 144 h postmortem and that the time groups were separated from each other on the PCA score plot. The prediction model showed high recognition capability (95.93%) and cross‐validation (83.72%). Our work suggests that MALDI‐TOF MS can be used to determine the PMI.

Identification of pulmonary edema in forensic autopsy cases of sudden cardiac death using Fourier transform infrared microspectroscopy: a pilot study

Many studies have proven the usefulness of biofluid-based infrared spectroscopy in the clinical domain for diagnosis and monitoring the progression of diseases. Here we present a state-of-the-art study in the forensic field that employed Fourier transform infrared microspectroscopy for postmortem diagnosis of sudden cardiac death (SCD) by in situ biochemical investigation of alveolar edema fluid in lung tissue sections. The results of amide-related spectral absorbance analysis demonstrated that the pulmonary edema fluid of the SCD group was richer in protein components than that of the neurologic catastrophe (NC) and lethal multiple injuries (LMI) groups. The complementary results of unsupervised principle component analysis (PCA) and genetic algorithm-guided partial least-squares discriminant analysis (GA-PLS-DA) further indicated different global spectral band patterns of pulmonary edema fluids between these three groups. Ultimately, a random forest (RF) classification model for postmortem diagnosis of SCD was built and achieved good sensitivity and specificity scores of 97.3% and 95.5%, respectively. Classification predictions of unknown pulmonary edema fluid collected from 16 cases were also performed by the model, resulting in 100% correct discrimination. This pilot study demonstrates that FTIR microspectroscopy in combination with chemometrics has the potential to be an effective aid for postmortem diagnosis of SCD.